Activated T-cell express CD25, the p55 chain of the IL-2 receptor. Here we report a reliable procedure for rapid determination of human gamma delta T cell activation by microfluorimetric measurement of CD25. Three days after initiation of culture, CD25 expression was determined. The sensitivity of this detection method was compared with [3H]thymidine incorporation at day 8. This proliferation assay allowed 3-5-fold higher dilution of the stimulatory ligand. However, monitoring of CD25 expression speeded up screening by 5 days. Therefore, for rapid screening of gamma delta T cell stimulation, e.g. for identification of activating ligands, monitoring of CD25 at day 3 is superior to [3H]thymidine measurement.